Clinical two‐photon microendoscopy

König, Karsten; Ehlers, Alexander; Riemann, Iris; Schenkl, Selma; Bückle, Rainer; Kaatz, Martin

doi:10.1002/jemt.20445

Cited by 152 publications

(142 citation statements)

References 11 publications

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“…The ageing skin of young and old people show a significant difference with the semi-quantitative evaluation of dermal collagen and elastin fibre network assessed in vivo by the multiphoton excitation imaging (Koehler et al, 2006). Procedures of selective imaging as well as spectral fluorescence lifetime analysis by means of multiphoton laser tomography support diagnostic decisions and may improve the process of non-invasive early detection of melanoma (König et al, 2007;Dimitrow et al, 2009) (Fig. 9) and non-melanoma skin cancer (Paoli et al, 2008).…”

Section: In Vivo Drug Screening and Administration In Skin And Kidneymentioning

confidence: 99%

Two‐photon microscopy of deep intravital tissues and its merits in clinical research

Wang

König

2010

Journal of Microscopy

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177

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SummaryMultiphoton excitation laser scanning microscopy, relying on the simultaneous absorption of two or more photons by a molecule, is one of the most exciting recent developments in biomedical imaging. Thanks to its superior imaging capability of deeper tissue penetration and efficient light detection, this system becomes more and more an inspiring tool for intravital bulk tissue imaging. Two-photon excitation microscopy including 2-photon fluorescence and second harmonic generated signal microscopy is the most common multiphoton microscopic application. In the present review we take diverse ocular tissues as intravital samples to demonstrate the advantages of this approach. Experiments with registration of intracellular 2-photon fluorescence and extracellular collagen second harmonic generated signal microscopy in native ocular tissues are focused. Data show that the in-tandem combination of 2-photon fluorescence and second harmonic generated signal microscopy as two-modality microscopy allows for in situ co-localization imaging of various microstructural components in the whole-mount deep intravital tissues. New applications and recent developments of this high technology in clinical studies such as 2-photon-controlled drug release, in vivo drug screening and administration in skin and kidney, as well as its uses in tumourous tissues such as melanoma and glioma, in diseased lung, brain and heart are additionally reviewed. Intrinsic emission two-modal 2-photon microscopy/tomography, acting as an efficient and sensitive non-injurious imaging approach featured by high contrast and subcellular spatial resolution, has been proved to be a promising tool for intravital deep tissue imaging and clinical studies. Given the level of its performance, we believe Correspondence to B.-G. Wang. Tel: +49 3641 938496; fax: +49 3641 938552; e-mail: Baogui.Wang@mti.uni-jena.de that the non-linear optical imaging technique has tremendous potentials to find more applications in biomedical fundamental and clinical research in the near future.

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Section: In Vivo Drug Screening and Administration In Skin And Kidneymentioning

confidence: 99%

Two‐photon microscopy of deep intravital tissues and its merits in clinical research

Wang

König

2010

Journal of Microscopy

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177

131

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“…This limitation may be overcome by technical developments of multi-photon microscopy scanner probes connected to optic fibres, which have recently facilitated in vivo imaging of the mouse central nervous system over extended periods of time [10,11,18]. König et al have further demonstrated that multi-photon microscopy can be performed through optical rods [12]. However, the device used for this study does allow fast optical serial biopsies during the operation.…”

Section: Discussionmentioning

confidence: 99%

Multi-photon excitation fluorescence microscopy of brain-tumour tissue and analysis of cell density

Kantelhardt

Leppert

Kantelhardt

et al. 2009

Acta Neurochir (Wien)

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Background Recent studies have highlighted the importance of the complete resection of a brain tumour but the task often remains a challenge for the neurosurgeon. New technologies which add objective information beyond visualisation provided by the traditional operating microscope are required. In this study, we have analysed the cellular density of the tumour/brain interface using three dimensional multi-photon microscopy intensity-images of experimental gliomas and human brain-tumour biopsy samples. Methods The density of cellular nuclei was determined in specimens of experimental gliomas in a mouse model and human brain tumour biopsies by analysis of optical tissue sections. Three dimensional multi-photon microscopy image stacks were compared to serial H&E stained sections of conventional histolopathology. Findings Both techniques consistently showed a good correlation of cell density values in solid tumour tissue of experimental gliomas versus adjacent brain. The multiphoton microscopy analysis of human biopsy specimens showed that optical analysis of native tissue provided information on the cellular density. Conclusions Multi-photon microscopy is an efficient and rapid tool for the study of brain and brain tumour tissue. Multi-photon microscopy allows the detection of individual tumour cells and tumour cell clusters in native tissue biopsies and may therefore provide a tool in the identification of highly cellular lesions during the resection of brain tumours.

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“…GRIN rod lens can also be used directly to couple with a microscope objective with similar or matching numerical aperture for microendoscopic imaging. In this case the pitch length of the GRIN lens to be used can be slightly shorter than the integral multiple of ½ pitches [21]. The practical issues of using a single GRIN lens are 1) the physical length of the GRIN lens that can be made useful for deep tissue probing, 2) the diameter of the GRIN lens which is desired to be small for microendoscopic applications and 3) the optical artifacts generated by the GRIN lens (e.g.…”

Section: Laser Scanning Microendoscopy Using Grin Lens Probesmentioning

confidence: 99%

“…GRIN lens is convenient for coupling light into fiber optics due to its cylindrical shape and therefore widely used as a component in telecommunication devices. As a lens, it also finds applications in the areas of optical spectroscopy [10,11] and imaging including optical coherent tomography (OCT) [12,13], endoscopy [14][15][16] and optical microscopy [17][18][19][20][21][22]. GRIN lenses were reported to be implemented in conjunction with both confocal and two-photon fluorescence microscopes for in vivo imaging of the brain [18,19,22].…”

Section: Introductionmentioning

confidence: 99%

“…To date, microscopic or microendoscopic applications using GRIN lens systems were mostly found in imaging of the brain in live rodents [18,19,22]. Recently, an application of using GRIN lens for dermatological inspection of volunteer patients was proposed [21]. We are interested in using a GRIN lens system to extend its microscopic application for intravital deep kidney imaging.…”

Section: Introductionmentioning

confidence: 99%

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Deep tissue microscopic imaging of the kidney with a gradient-index lens system

2008

Optics Communications

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Intravital microscopy using two-photon excitation is proven to be a valuable tool for studying the kidney and associated disease processes. However, routine performance of intravital kidney imaging is limited by the fact that fluorescence signal is attenuated by the tissue and at certain tissue depth lost its strength completely. For most of the animal tissues, this finite imaging depth is limited to a few hundred microns. Currently it is not possible to non-invasively image the kidney beyond the superficial tissue layers of the cortex. This has imposed significant limitations on the animal models one can use for imaging since structure such the glomerulus is typically located below the superficial layer of the cortex that can not be imaged using a conventional fluorescence microscope. Here we report the use of a needle-like lens system based on gradient-index (GRIN) microlenses capable of transferring high quality fluorescence images of the tissue through a regular microscope objective for deep tissue imaging of the kidney. By combining this GRIN lens system with a Zeiss LSM 510 NLO microscope, we are able to extend the imaging depth for kidney tissues far beyond the few hundred microns limit. This GRIN lens imaging system provides an alternative microendoscopic imaging tool that will enhance current intravital kidney imaging techniques for studying structural and functional properties of local tissues at locations below the superficial layers of the kidney.

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Clinical two‐photon microendoscopy

Cited by 152 publications

References 11 publications

Two‐photon microscopy of deep intravital tissues and its merits in clinical research

Two‐photon microscopy of deep intravital tissues and its merits in clinical research

Multi-photon excitation fluorescence microscopy of brain-tumour tissue and analysis of cell density

Deep tissue microscopic imaging of the kidney with a gradient-index lens system

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